EP2322914B1 - Vorrichtungen und verfahren zum nachweis von analytkonzentrationen - Google Patents
Vorrichtungen und verfahren zum nachweis von analytkonzentrationen Download PDFInfo
- Publication number
- EP2322914B1 EP2322914B1 EP10180848.3A EP10180848A EP2322914B1 EP 2322914 B1 EP2322914 B1 EP 2322914B1 EP 10180848 A EP10180848 A EP 10180848A EP 2322914 B1 EP2322914 B1 EP 2322914B1
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- EP
- European Patent Office
- Prior art keywords
- component
- reagent
- arrangement
- analyte
- detectors
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Classifications
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Definitions
- the present invention is directed to techniques and devices for detection of the presence and/or concentration of an analyte and to techniques and devices for the effective collection, transportation and quantization of small quantities of body fluids.
- US 6, 312, 888 B1 disclose an article for the concentration of an analyte e.g. glucose in blood.
- the invention involves an article comprising a multiple layer effect.
- the state of the art has been advanced through the provision of devices and techniques such as those described further herein, for accurately, efficiently, and economically determining the presence and/or concentration of an analyte. According to the present invention, the state of the art has been advanced, especially, but not exclusively, within the context of personal glucose monitoring devices and techniques.
- the present invention provides an arrangement as set forth in claim 1.
- CMOS or CCD sensors may include any suitable sensor.
- the present invention is thus not limited to embodiments of the invention including CMOS or CCD sensors, photodiodes, infrared, fluorescent, ultraviolet or electrochemical sensors.
- the present invention provides an assay pad construction comprising: a first component comprising a constituent to separate red blood cells from plasma and further comprising a diffuse reflective material constituent; a second component comprising a chemical reagent; and a third component comprising a polyamide-containing mesh.
- the present invention provides an assay pad comprising: a first component comprising a diffuse reflective material; a second component comprising a hydrophilic material; a third component comprising a reagent; and a fourth component comprising a mesh or a membrane.
- first, second, third and fourth components does not limit the present invention to embodiments where each of these components is physically separable from one another.
- a single physical element of the invention may perform the features of more than one of the claimed first, second, third or fourth components.
- a plurality of separate physical elements working together may perform the claimed features of one of the claimed first, second, third or fourth components.
- the present invention provides a method as set forth in claim 6.
- Figure 1 is one such arrangement 100 for the optical detection of the presence and/or concentration of an analyte such as glucose.
- the arrangement 100 can generally be described as a diffuse transmission arrangement.
- the substance to be analyzed is transported via a conduit 12.
- the conduit 12 can be in the form of a hollow member or needle or any other skin piercing element, including, but not limited to lancets.
- the needle can be of a very narrow gage, or a so-called microneedle.
- Such a needle typically having a size on the order of 40-100 micrometers. It is to be understood, however, that diameters of less than 40 micrometers, or more than 100 micrometers are also contemplated within the scope of the present invention.
- the microneedle may also act as a skin penetration member as well as a conduit.
- the skin penetration member may be in the form of a solid lancet (not shown), which acts to produce a sample of bodily fluid which is then transported via conduit, e.g.-12.
- the substance to be analyzed is transported via the conduit 12 to a chamber 14.
- the chamber may have any suitable form.
- the chamber 14 is formed by a lower member 16 and an upper member 18. These members 16, 18 can be constructed to allow light to be transmitted therethrough.
- An assay pad 20 is provided within the chamber 14.
- the pad 20 receives the substance to be analyzed.
- the pad 20 is provided with a substance that reacts with the analyte. The results of this reaction is detectable and the data generated through detection is used to determine the presence and/or concentration of the analyte.
- the pad 20 is provided with a construction to achieve this objective, and others. A discussion of the specifics of the pad 20 will be deferred.
- a light source 22 can be provided so as to produce light that is incident upon a first side of the pad 20. Any suitable light source is envisioned. According to a preferred embodiment, the light source comprises a light emitting diode (LED). The LED or light source 22 can provide incident light of any suitable wavelength. Wavelengths on the order of 500 - 700nm are suitable. For example, incident light with a wavelength on the order of 670nm may be transmitted from light source 22. At approximately 670nm, absorption of the incident light by hemoglobin tends to be reduced.
- LED light emitting diode
- An optical detector 24 is positioned so as to receive light transmitted through a second side of the pad 20, the second side being opposite the first side.
- the optical detector can be a digital imaging device, such as a complementary metal oxide semiconductor (CMOS) device, a charge couple device (CCD), or any photodiode or any other suitable detector, including, but not limited to infrared sensors, including near and mid field infrared sensors. Electrochemical and reagent-less sensing techniques are also contemplated within the scope of the resent invention.
- the imaging device may comprise an array of individual detectors.
- the implementation of digital imaging devices permits the analysis of samples that are not of uniform volume. This is accomplished at least in part by the ability of individual detectors to take readings, which are then averaged or combined to generate an overall reading. The larger the sample, the larger the "spot" formed on the pad 20, and the more individual detectors pick up a reading of the reaction occurring in the pad 20.
- the assay of the present invention is not dependent upon the volume of the sample being analyzed. When an assay is incorporated into a monitoring device wherein the monitoring is performed in an automated fashion, the user is often unable to determine if a sample of body fluid of a particular volume has been obtained.
- the accuracy of the assay is directly linked to a specific volume of sample undergoing analysis (e.g. - "microtitration ").
- the present invention breaks this dependence, thereby providing robust and reliable devices, such as personal glucose monitoring devices, that can function effectively over a range of potential sample volumes.
- Other details of the detector 24, and associated components, will be deferred to later text.
- the arrangement 100 may be incorporated into a larger overall device. According to one embodiment, the arrangement 100 is incorporated into a personal glucose-monitoring device. According to one alternative, the device is ambulatory. For example, the device is wearable by a user and autonomous in its operation. The arrangement 100 can be incorporated into devices, such as the ones described in U.S. Patent Publication No. US 2002/0087056 or US 2003/0153900 .
- FIG. 2 An alternative arrangement 200 is depicted in Figure 2 .
- the arrangement 200 can generally be described as a diffuse reflection arrangement.
- the light source 22 is provided such that it transmits light which is incident upon a first side of the assay pad 20. At least a portion of this incident light is reflected off of the first side. This reflected light can then be collected by the optical detector 24, which is disposed on the same side as the light source 22, relative to the test pad 20.
- the light source 22 and the optical detector 24 are disposed, relative to the reaction chamber, on a side which is opposite from the side of the reaction chamber into which the conduit 12 introduces the substance to be analyzed. This orientation is shown in Figure 2 .
- the test pad 20 may be provided with a blocking layer 21.
- a blocking layer 21 can be formed of any suitable material, such as zirconium oxide or titanium oxide.
- the reading taken by the detector 24 is insensitive to the contents of the chamber 14 since the contents of the chamber 14 are not in direct communication with the side of the test pad 20 being read.
- FIG. 3 Another alternative arrangement constructed according to the principles of the present invention is illustrated in Figure 3 .
- the arrangement 300 can generally be described as an edge or waveguide illumination arrangement.
- a light source 22 is arranged such that a diverging light beam strikes an edge of a transparent waveguide member 32, and couples the transmitted light inside the waveguide, as illustrated by the broken lines contained in Figure 3 .
- ⁇ C sin -1 (n 2 /n 1 ), where ⁇ C is the angle of incidence that will cause incident light to become trapped in the waveguide, and n 2 and n 1 are the indices of refraction for a first material and a second material that define a boundary across which the light is attempting to travel (e.g. - air and test strip material).
- the coupling edge of the waveguide may be treated with an anti reflective coating or an index-matching medium.
- an anti reflective coating or an index-matching medium For example, one or more thin layers of silicon oxide may be applied to the coupling edge. A thickness of approximately 200nm is an exemplary thickness.
- TIR total internal reflection
- the assay pad 20 is provided with a surface in communication with the reflected light inside the waveguide, which is light scattering.
- Light impinging on this surface 34 of a test pad 20 is scattered at all angles. As illustrated by the dotted lines in Figure 3 , some of the light is scattered at a large angle relative to the normal of surface 34. This light is also trapped inside the waveguide. However, some of the light is scattered at a relatively small angle compared to the normal of surface 34, as illustrated by the solid double arrow lines of Figure 3 , and is consequently transmitted outside of the waveguide and received by the detector 24.
- FIG. 4 Another arrangement 400 of the type described above, is also illustrated in Figure 4 .
- arrangements of this type 300, 400 are such that light provided by the light source 22 is incident upon the waveguide at a relatively small angle to the surface thereof, a larger amount of this incident light is transmitted through the waveguide, when compared with arrangements wherein the incident light is provided at a relatively steep angle to the incident surface.
- the light source is an LED-type source, less power is required in order to provide the necessary amount of incident light to perform the assay, and power consumption can be minimized.
- the light source 22 can be closely aligned with the edge of the waveguide 32, and thus may provide for a compact configuration.
- the detector element 24 can be positioned in close proximity to the waveguide 32, assay pad 20 and light source 22 ( Figure 4 ), thereby enabling it to be more protected from stray light sources.
- an imaging lens 52 is provided and disposed such that it captures and focuses light reflected from the assay pad 20 toward the optical detector 24.
- the lens 52 may comprise a refractive or defractive element (Fresnel lens).
- the lens 52 is utilized in connection with an edge illumination-type arrangement.
- the lens 52 is illustrated as being disposed at some distance from the waveguide 32 which forms the upper portion of the reaction chamber 14, the lens member may also be integrated into either the surface of the detector 24, or into the upper member (e.g. - 18, Figure 1 ) or waveguide 32.
- Figure 6 depicts an alternative arrangement 600 which comprises a diffuse reflection type arrangement which incorporates an imaging lens member 52, as described above.
- the arrangement 700 includes an edge-illumination type arrangement including waveguide 32.
- An imaging lens 54 is also included. In this arrangement, the imaging lens is integrated into either the waveguide 32 or the detector 24.
- Figures 8a and 8b illustrate in further detail certain aspects of an additional embodiment of the present invention.
- Figures 8a and 8b illustrate and arrangement 80 which can be generally described as a vertical-flow assay.
- the arrangement 800 includes a channel member 82 having an open "bottom", an assay pad 20 in communication with the open bottom of channel member 82, and an optical detector, preferably in the form of an array 84 of digital imaging devices 85.
- lens elements 86 are incorporated into the arrangement. Although lens elements 86 are illustrated as being in the form of discrete individual lenses in close proximity to the individual detector elements or pixels 85, it is within the scope of the present invention that alternative constructions are possible. For example, the lenses 86 could be spaced from the detectors 85, and/or a single integrated lens construction could be utilized instead of discrete lenses 86.
- the channel member 82 is elongated.
- the substance being analyzed such as a sample of body fluid is introduced at one end 81 of the channel member 82 and flows laterally or axially therethrough.
- the substance being analyzed then permeates the test pad 20 disposed thereunder.
- the open channel member 82 defines a particular volume, it is not necessary to completely fill the channel member 82 in order to conduct an accurate analysis.
- a particular volume of substance to be tested may only partially fill channel member 82, for example, it may only fill channel member 82 up to the point defined along line 83.
- the independent detector pixels 85 of the array 84 on the left-hand side of line 83 are activated and will take a reading from the analysis side of the test pad 20 once the analyte present in substance under analysis reacts with the chemicals contained in the test pad 20. Those individual detector elements of the array which are on the left-hand side of line 83 will generate a signal which is indicative of the presence of this reaction. Those individual detector elements 85 of array 84 which are on the right hand side of line 83 will not generate such a reading. Thus, the readings or data generated by each individual detector element 85 of the array 84 is taken and analyzed, such as by averaging, to generate an overall reading of the presence and/or concentration of a particular analyte contained in the substance under analysis.
- the volume of the channel member 82 is known, and the length of the detector element contained in the array 84, as well as any correction due to the presence of lens elements 86 are known, the volume of the sample under analysis can be estimated based upon the number of pixels which detect the reaction between the analyte and the chemical reagents contained in test pad 20.
- the time between the readings taken by the detectors 85 of the array 84 can also be monitored by the electronics of the arrangement.
- the specific means of implementing this monitoring being well within the skill of those in the art.
- the arrangement of the present invention can also analyze the kinetics of the reaction between the analyte and the chemical reagents, thereby offering the possibly to provide additional useful information that can be used to provide an insightful analysis of the sample.
- An arrangement 80 of the type depicted in Figures 8a through 8b can advantageously be utilized to analyze very small volumes of a substance to be tested.
- the arrangement can be utilized to assay 1-500 nanoliter (nL) volumes.
- the substance to be tested may include whole blood, interstitial fluid, or combinations thereof.
- a sample of body fluid is introduced along direction B 1 from the left hand side 81 of channel number 82 through any suitable means, such as conduit 14 (see, e.g. - Figures 1 through 2 ).
- the sample is extended laterally into the elongated channel member 82.
- the volume of channel member 82 is 300 nanoliters.
- the sample is extended over a relatively long length, which can be on the order of a few millimeters.
- the channel is approximately 250 micrometers (width) x 140 micrometers (height) x 7,000 micrometers (length).
- a color change in the assay pad 20 may be produced by a chemical reaction triggered by the presence of a specific analyte contained in the sample of body fluid, such as glucose. This color change can be detected on the opposite of the assay pad from which the sample of body fluid is introduced.
- This color change is detected by a suitable detection device, such as the optical detector array 84, which may be composed of a plurality of digital imaging detectors 85.
- these digital detectors comprise either a CMOS, CCD, or photo diode array or infrared, ultraviolet or fluorescent sensors. It is to be understood, however, that other sensors may be used, all keeping within the scope of the present invention.
- red blood cells can be filtered or blocked from the opposite side of the test pad 20, which is generally viewed as being preferable so as to enable a more accurate reading of the levels of analyte present in the sample.
- the chemical reagents and dye products utilized in test pad 20 can be chosen from any number of well know substances. In any event, these chemistries should provide a discernable and readable reaction over an appropriate range of analyte concentration levels. In the case where the device is to be utilized to monitor the concentration of glucose contained in samples which comprise mainly whole blood, these chemistries should be able to produce reactions which can then be detected to indicate a glucose concentration ranging from, for example, 40-500 mg/dL.
- the detector array 84 is formed from a plurality of individual CMOS-type detectors 85.
- CMOS detectors are less costly than other types of digital imaging devices.
- various "clock" or time-driven electronic devices may be incorporated. Data can be taken from the detectors 85 and fed to such devices (directly or from storage) thereby enabling the monitoring and interpretation of the kinetics and other aspects of the reaction between the analyte and the reagent.
- uniformity and accuracy of the assay is also promoted. For example, readings are taken and utilized from multiple individual detector elements, or pixels, are less susceptible to variations in uniformity of the materials of the assay pad, or other localized abnormalities, which can be accounted for and corrected by the interpretation and manipulation of the data generated by the individual pixels 85 of the detector array 84. For example, statistical analyses and associated algorithms may be used to correct for such localized defects, and therefore improve the accuracy of the overall reading generated by the device.
- Figures 9 and 10 represent a scan performed by an arrangement 80 such as the one depicted in Figures 8a and 8b .
- Figure 9 is a picture of a test spot formed on a commercially-available assay pad.
- Figure 10 is a plot of the signal generated by an array of detector elements 84 as they are scanned along the dashed line contained in Figure 9 . As illustrated in Figure 10 , the lower voltage readings correspond to the area occupied by the test spot of Figure 9 along the dashed line.
- FIG 11 is a schematic illustration of one possible construction for the assay pad 20.
- the assay pad construction 1100 has a sample application side SA as well as an analysis side A.
- the assay pad 1100 is composed of at least 3 constituent components: a blocking component 1102; a membrane component 1104; and a chemical reagent component 1106.
- a blocking component 1102 When the sample to be analyzed is composed at least in part of whole blood, red blood cells are separated from the plasma, which is then transported to the reagent component 1106 which includes enzymatic chemistries selected to produce a chemical reaction with one or more analytes present in the sample.
- the blocking component 1102 may include an agent to filter or pre-filter red blood cells.
- the blocking component 1102 may include zirconium oxide (ZrO 2 ).
- the membrane component 1104 can be formed of any suitable material.
- the membrane may be a high light transmitting material, or optionally can be an opaque or substantially opaque material. Suitable materials may include styrene and/or nylon.
- the membrane component 1104 can be in woven or non-woven form. According to one aspect, the membrane is in the form of a woven mesh.
- the reagent component 1106 is chosen according to its ability to react with the analyte under investigation. Any suitable reagent chemistry can be utilized. Specific examples of suitable material include a glucose oxidase (e.g. - from Biocatalysts product G575P), a soybean peroxidase (e.g. - from Organic Technologies, product 73930.1 medical diagnostic grade), an amino antipyrine hydrochloride (e.g. - from TCI, product A0257) and an Aniline derivative dye for a "Trinder” reaction (see, e.g. - P. Trinder, "Determination of Glucose in Blood Using Glucose Oxidase with an Alternative Oxygen Acceptor").
- a glucose oxidase e.g. - from Biocatalysts product G575P
- soybean peroxidase e.g. - from Organic Technologies, product 73930.1 medical diagnostic grade
- the blocking component 1102 and/or the reagent component 1106 may be partially or entirely subsumed and contained within the membrane component 1104.
- a surface "layer" of the blocking component 1102 and/or reagent component 1106 may be present.
- the blocking component 1102 and/or the reagent component 1106 may be entirely subsumed, contained or impregnated within the membrane component 1104, such that no distinct blocking component "layer” or reagent component "layer” is present at all.
- the present invention encompasses embodiments where more than one of the claimed (first, second, third, fourth) components are performed by the same physical element. Conversely, the present invention also encompasses embodiments where a plurality of different physical elements work together to perform the features of one of the claimed (first, second, third, fourth) components. The present invention also encompasses embodiments with no sample pad at all.
- the blocking component 1102 and/or reagent component 1106 may be applied to the membrane component 1104 in any suitable manner.
- the blocking component 1102 and/or reagent component 1106 may be coated upon the membrane 1104.
- suitable coating techniques includes spin coating.
- Figure 12 illustrates one possible alternative construction 1200 for the assay pad 20.
- the assay pad construction 1200 is composed of at least 4 components: a prefilter and reflective component 1202, a membrane component 1204, a reagent component 1206, and a mesh component 1208.
- a prefilter and reflective component 1202 the red blood cells are separated from the plasma, so that components 1202 and/or 1204 retain the red blood cells without lysis.
- the plasma is then vertically transported to the reagent component 1206 which includes enzymatic chemistries.
- suitable materials include any suitable material, which are generally known in the art. Specific examples of suitable materials include a glucose oxidase (e.g.
- the component 1204 may comprise any suitable blood-filtering component or material.
- suitable blood-filtering component or material Some materials which have been used to separate red blood cells from plasma and samples of whole blood include glass fibers, mesh cloth, unwoven mats, asymmetric membranes, and porous polymers.
- component 1204 Regardless of what type of material is chosen for component 1204, the avoidance of spreading or lateral diffusion, which may occur with such materials as poly (ethersulfone) membranes, is desirable.
- An exemplary suitable material is, a hydrophilic poly(vinylidene fluoride) unwoven membrane. Such membranes are commercially available from the Millipore Corporation (e.g. - with 0.1 micron openings).
- Other suitable materials include unwoven membranes comprising a polyamide.
- Other materials may be incorporated to achieve various performance objectives, such as poly(acrylic acid) (PAA), hydroxyethyl cellulose, sodium alginate, and gelatin.
- the pad 1200 may further include a component 1202, such as a prefilter and/or diffuse reflection component.
- a component 1202 such as a prefilter and/or diffuse reflection component.
- the component 1202 when included, serves to improve separation of red blood cells from a sample containing whole blood, and/or increase the reflection of incident light upon the opposite side of the pad when undergoing optical analysis.
- Suitable materials for use as component 1202 include a zirconium oxide (ZrO 2 ), optionally combined with a polymer binder material.
- the component 1202 can be applied to the membrane, or component 1204, by using any suitable technique.
- One such suitable technique is spin coating. Spin coating promotes uniform distribution of the component 1202 onto and/or within the membrane component 1204.
- the component 1202 may be partially, or entirely contained within the membrane component 1204, thus effectively combining these two components into a single unitary member, rather than the illustrated distinct layers.
- the component 1202 comprises a prefilter and diffuse reflection material and a polymer binder
- the presence of the polymer binder tends to form at least a thin layer or film over the membrane component 1204.
- This thin film may have any suitable thickness, for example, the component 1202 may partially permeate the membrane component 1204 and leave a thin film having a thickness on the order of less than 5 micrometers on top of the membrane component 1204.
- the component 1206 comprises a reagent which may be formed from any suitable composition, such as the chemicals described above, and may optionally be provided in a binder of poly(acrylic acid), hydroxypropylmethylcellulose (HPMC) or similar polymers that reduce lateral spreading of the substrate or sample undergoing analysis.
- a reagent which may be formed from any suitable composition, such as the chemicals described above, and may optionally be provided in a binder of poly(acrylic acid), hydroxypropylmethylcellulose (HPMC) or similar polymers that reduce lateral spreading of the substrate or sample undergoing analysis.
- the mesh component 1208 and the reagent 1206 component may also be integrated into a single layer or constituent element of the test pad 20.
- reagent material 1206 can be coated onto the mesh component 1208.
- Any suitable coating technique may be utilized.
- a spin coating technique is one such technique for providing the reagent on the mesh component 1208 which allows for precise and relatively thin coating of the reagent component 1206 to be applied to the mesh component 1208. According to one construction, this "coating" is entirely absorbed and contained within mesh component 1208.
- the mesh component 1208 should be permeable to oxygen in order to promote the reaction of the analyte with the chemical reagent component 1206.
- the mesh component 1208 can be formed from a high reflectance woven mesh, such as nylon.
- mesh component 1208 is formed from an open polyamide or nylon mesh material.
- This nylon mesh material having a reagent incorporated therein is believed to increase the sensitivity of the assay.
- the open structure of the nylon mesh in conjunction with the controlled amount of the reagent component material 1206 incorporated therein, increases the diffuse reflection of light incident thereon. This increased diffuse reflection is believed to be caused by the multiple reflections and increased path lengths provided by the open nylon mesh structure.
- the mesh component 1208 has openings have a dimension which is on the order of 68 microns, and a filament diameter which is on the order of approximately 50 microns.
- Circle A 1 illustrates the absence of reagent material within the mesh, wherein the reagent material has been consumed by chemical reaction with the analyte.
- Circle A 2 indicates an area of the mesh in which the reagent material is still disposed thereon.
- the cylindrical fibers of the nylon mesh may scatter the light onto the external exposed layer or surface thereby introducing multiple reflection and increased path links.
- the scattering of incident light is largely dependent upon a refractive index of the cylinders which form the filaments of the mesh.
- the refractive index of nylon is relative large, on the order of 1.53, and this is beneficial.
- the above-described application of the reagent component 1206 to the mesh component 1208 can simplify manufacture.
- test pad construction 1400 includes three basic components: a filter and/or diffuse reflective component 1402, a chemical reagent component 1404, and a membrane component 1406.
- a sample to be analyzed is applied to the sample application side SA of test pad 1400.
- red blood cells are separated from the plasma by component 1402 and are transported to the reagent component 1404, which may be partially or entirely impregnated it the membrane component 1406.
- the component 1402 may be formed from any suitable construction that provides filtration and/or diffuse reflection functions.
- suitable constructions have been described above.
- the component 1402 may comprise a zirconium oxide material combined with a permeable polymer binder material, such as HPMC, that limits lateral diffusion.
- a zirconium oxide material combined with a permeable polymer binder such as vinyl butyral. The zirconium oxide and polymer binder are combined in any suitable proportions, such as 20:80.
- the reagent component 1404 includes at least one chemical reagent chosen to provide a suitable reaction with the analyte(s) under investigation.
- Any suitable reagent can be chosen.
- the reagent may comprise a glucose oxidase, soy bean peroxidase, amino antipyrine hydrochloride, and/or aniline derivative dyes.
- the chemical reagent may optionally be combined with a binder component.
- the binder component is preferably formed from a material which reduces the tendency for lateral spreading of the sample within the test pad 1400. Suitable examples binder material examples include HPMC and PAA.
- a suitable membrane component 1406 can also be provided.
- the membrane component 1406 can comprise a fibrous unwoven material.
- One specific example includes a cellulose acetate unwoven fibrous membrane material.
- Both the filter/diffuse reflective component 1402, as well as the reagent component 1404 can be applied to the membrane component 1406 by any suitable technique.
- the reagent component 1404 is applied to the membrane component 1406 by a coating technique, such as spin coating.
- Reagent component 1404 may be partially, or entirely impregnated within the membrane component 1406.
- a thin film or layer of reagent component 1404 may be present on the top surface of the membrane component 1406 (i.e. - the side closest to the sample application side SA of the test pad 1400). When present, this thin top layer can be on the order of a few microns in thickness.
- the reagent component may be entirely subsumed, contained or impregnated within the membrane component 1406, such that no distinct layer is present on the top surface of the membrane component 1406 (i.e. - the surface of membrane 1406 which is closest to the surface application side SA of the pad 1400).
- the filter/diffuse reflective component 1402 can then be applied subsequent to application of the reagent component 1404.
- the component 1402 can also be applied by any suitable technique, such as spin coating.
- a filter/diffuse reflective component comprising a zirconium oxide and hydrophobic permeable polymer binder material is spin coated at approximately 1000 rpm onto the membrane/reagent components 1406/1404.
- the filter/diffuse reflective component 1402 may be partially impregnated within the membrane 1406, leaving a thin film layer at the surface application side SA of the combined membrane and reagent components 1406 and 1404. When present, this thin surface layer can be a few microns in thickness, for example, on the order of less than 5 microns in thickness.
- the filter/diffuse reflective component 1402 may be entirely subsumed, contained, or impregnated within the combined reagent and membrane components 1404 and 1406.
- test pad construction 1500 is comprised of at least four basic components: a filter/diffuse reflective component 1502, a permeable material component 1504, a reagent component 1506, and a membrane component 1508.
- the general functionality of the test pad 1500 and the functionality of the individual components thereof are similar to that previously described.
- the component 1502 may comprise any suitable filter and/or diffuse reflective material.
- the component 1502 may comprise a zirconium oxide material with or without a polymer binder.
- the polymer binder is preferably a permeable material.
- suitable permeable polymer binders include HPMC and an acrylic resin, such as Elvacite®.
- the component 1504 is preferably comprised of a suitable permeable material which can be hydrophobic.
- the hydrophobic material can comprise any suitable form.
- the component 1504 may comprise a film of a hydrophobic polymeric material, such as PAA.
- the film can be formed by any suitable technique.
- a film of PAA is formed by a fast evaporating solvent/PAA mixture.
- the mixture may contain 10% by weight (relative to the weight of the solvent) of PAA.
- the component 1506 may comprise a suitable reagent material.
- the reagent material may be combined with a binder.
- the reagent component may be chosen from any suitable substance or combination of substances, as described above.
- the binder material comprises a permeable hydrophobic binder material.
- HPMC HPMC.
- the membrane component 1508 can be formed from any suitable material. According to one possible construction, the membrane component 1508 is formed from an unwoven fibrous mat.
- the reagent component 1506 can be spin coated upon the membrane component 1508. As discussed above, the reagent component 1506 may be partially, or entirely, impregnated within the membrane component 1508. Thus, the reagent component 1506 may or may not comprise a thin surface layer formed on top of the membrane component 1508 (the top side being closest to the surface application side SA of the test pad 1500).
- the hydrophobic material component 1504 can then be applied.
- the hydrophobic material can be formed as a slurry or mixture with a fast evaporating solvent, such as methanol.
- the mixture is then coated onto the combined membrane and reagent components 1508 and 1506. Subsequent to evaporation, a thin film of the hydrophobic material is left on top of the combined reagent/membrane component.
- the component 1502 may then be applied to the previously described combined components.
- a combination of filter/diffuse reflective material, e.g. - ZrO 2 , and a polymer binder material, such as the ones described above may be combined in the form of a slurry or mixture and then coated upon the previously combined components.
- a slurry containing up to 10% by weight of ZrO 2 may be formed and then spin coated onto the previous combined constituent components 1504, 1506, and 1508.
- the component 1502 may be partially or entirely impregnated or subsumed within the previously combined components.
- the component 1502 may be indistinguishable from the previously combined components or, may form a thin film or layer on top of the previously combined components.
- test pad construction 1600 formed according to the principles of the present invention is illustrated in Figure 16 .
- the test pad construction 1600 illustrated therein is formed from at least three basic constituent components: a filter/diffuse reflective component 1602, a permeable hydrophobic material component 1604 and a combined reagent/mesh component 1606.
- the basic functionality of the individual as well as combined constituent components of the test pad construction 1600 is similar to that previously discussed.
- the filter/diffuse reflective component 1602 can be formed from any suitable material, and comprise any suitable construction.
- the component 1602 comprises a zirconium oxide material, which may optionally be combined with a binder material.
- the binder material is preferably a permeable hydrophobic binder material such as an acrylic resin. Such material is being commercially available, e.g. - sold as Elvacite®.
- the permeable hydrophobic material component 1604 can be comprised of any suitable material, and take any suitable form.
- the permeable hydrophobic material can comprise a film or layer.
- the permeable hydrophobic material may comprise PAA.
- the combined reagent/mesh component 1606, as implied is preferably the combination of two distinct parts or constituents. Namely, a reagent material such as those previously described, as well as a mesh material constituent. Preferably, these constituents are combined into a single, integral component 1606.
- the mesh material can be formed of any suitable material and take any suitable form.
- the mesh material can be formed from a woven polyamide material, such as a woven nylon mesh.
- the mesh may have openings of any suitable dimensions, such as either 30 microns openings or 41 micron openings.
- the reagent and the mesh component are combined or integrated by soaking the mesh material in a slurry formed from the reagent material.
- a slurry or mixture can be formed which comprises 20% by weight HPMC binder material plus a suitable reagent chemistry.
- the mesh material is soaked in this liquid slurry.
- the mesh material is then removed and dried, thereby leaving an integrated mesh/reagent component 1606.
- the hydrophobic component 1604 can be similarly be applied by any suitable technique.
- the hydrophobic material can be applied as a film formed from a fast evaporating solvent, like methanol.
- a hydrophobic material such as PAA is combined with a solvent to form a slurry.
- the slurry may comprise 10% by weight of the PAA material.
- the solvent is then applied to the combined reagent/mesh component 1606. Upon evaporation of the solvent, a thin film or layer of the hydrophobic material is left behind, thereby forming the hydrophobic material component 1604.
- the filter/diffuse reflective component 1602 may similarly be applied by any suitable technique, such as those previously described.
- a zirconium oxide material is added to a hydrophobic binder to form a slurry in an evaporative solvent such as toluene.
- the slurry contains approximately 10% by weight of acrylic resin such as Elvacite®.
- This slurry is then coated onto the combined hydrophobic and reagent/mesh components 1604 and 1606.
- Any suitable coating technique can be utilized, such as the aforementioned spin coating technique.
- the filter/diffuse reflective component 1602 is provided in the form of a thin layer or film on top of the combined hydrophobic material and reagent/mesh component 1604 and 1606.
- a further aspect of the present invention provides effective and efficient constructions and techniques that allow for the reliable collection and transport of relatively small quantities of body fluids, such as blood.
- body fluid includes at least the following components: blood, interstitial fluid, and combinations thereof.
- skin penetration member includes hollow needles and solid lancets, as well as multiple component constructions, such as concentric needles or a concentric hollow tubular collection member and needle arrangement.
- needle includes, but is not necessarily limited to, so-called “microneedles” or small gage needles.
- microneedles can have an outer diameter on the order of 40 - 200 ⁇ m and an inside diameter on the order of 25 - 160 ⁇ m.
- these "microneedles” can be of a small gage, such as on the order of 34 or 36 gage needle.
- sample of body fluid can be obtained from either within the wound beneath the surface of the skin and/or off the surface of the skin itself.
- Figures 17 and 18 are illustrative of an arrangement 2100 for the collection an analysis of a sample of body fluid 2108, such as blood.
- the illustrated arrangement 2100 generally comprises a skin-penetration member 2102, an analysis chamber 2104, an analysis component 2106, and an inner bore 2110 of the skin-penetration member 2102.
- body fluid 2108 can be drawn up through the inner bore 2110 in the direction of the heavy arrow via capillary forces.
- the embodiments of the present invention illustrated in Figures 17 to 23 operate in the same manner as described above with reference to Figures 1 to 16 .
- the analysis chamber 2104 is vented in order to facilitate capillary flow and preventing the build-up of a back pressure within the analysis chamber 2104 that would impede flow of the body fluid 2108.
- the inner bore 2110 can be modified in a manner that promotes capillary action.
- the inner bore 2110 can be coated with a substance that promotes capillary action.
- Figure 19 illustrates an arrangement similar to that previously illustrated in Figures 17 and 18 , however, the arrangement 2100 of Figure 19 does not provide a continuous uninterrupted path from the point of body fluid acquisition to the analysis chamber 2104. Instead, the arrangement 2100 of Figure 19 includes a capillary break at 2112.
- the arrangement depicted in Figure 20 also includes a capillary break 2114, but the capillary break 2114 has a construction designed to minimize the adverse effects on the capillary flow of body fluid 2108.
- the arrangement of Figure 20 is considered an improvement, at least in this respect, over the arrangement of Figure 19 .
- Figure 21 illustrated another arrangement 2100 constructed according to the principles of the present invention.
- the primary distinction between the arrangement of Figure 21 and the previously described arrangements is the use of an applied vacuum via a vacuum generation device 2116 in order to assist body fluid collection and transportation.
- the inner bore 2110 of the illustrated embodiment provides a continuous and uninterrupted path from the point of body fluid acquisition to the analysis chamber 2104.
- the vacuum generating device facilitates the flow of body fluid along this uninterrupted path.
- the application of a vacuum can also be beneficial in arrangements such as those illustrated in Figures 19 and 20 that have a capillary break (e.g. - 2112, 2114).
- FIG. 22 The arrangement depicted in Figure 22 is similar to the previous arrangements except that the analysis chamber 2104 is static, while the skin penetration member 2102 is moveable along the direction indicated at double-headed arrow 2118 relative thereto. This construction necessitates a moveable seal between the skin penetration member 2102 and the analysis chamber 2104 designed to maintain the capillary flow.
- the skin-penetration member 2102 and the analysis chamber 2104 are relatively fixed, as illustrated in Figure 23 . Although these two members are fixed relative to one another, and thus move together upon insertion of the skin-penetration member, the members can be moveable via connection to a hinge or translation stage (not shown).
- the present invention provides arrangements for the detection of the presence and/or concentration of an analyte in a sample of bodily fluid include diffuse transmission, diffuse reflection and edge or waveguide illumination arrangements.
- a vertical flow assay arrangement and/or technique is also disclosed, and includes a detection component that can be in the form of an array of optical detection elements.
- a number of assay pad constructions are described which may include at least one or more of the following components: a prefilter component, a reflective component, a membrane component, a reagent component, a mesh component, and a component to prevent lateral spreading. It is to be understood that in alternate embodiments, the present invention can be operated without an assay pad.
- the present invention also provides: a system for effective and efficient techniques and constructions that allow for quantitative measurement of the concentration of an analyte using algorithms.
- algorithms may be based on peak average, peak area, or peak value with respect to the measurement of the concentration of glucose levels present in a nanoliter-sized sample of body fluid using a CMOS-based detector device.
- the algorithms are constructed to select data from the measurement that results in improved assay statistics.
- non CMOS-based detectors may also be used, all keeping within the scope of the present invention.
- CMOS data three algorithms can be utilized for quantitative glucose measurements using CMOS data.
- glucose measurements were made in a diffuse reflection mode where a narrow wavelength LED emits light onto an assay device and signal is recorded with a CMOS array.
- CMOS complementary metal-oxide-semiconductor
- a color develops on the assay device and the diffuse reflectance signal detected by CMOS is reduced by the absorption of light. The level of reduction depends upon the glucose concentration.
- Figure 24 shows typical curves for CMOS detection.
- the background line shows the CMOS signal before glucose was delivered to the assay device.
- the glucose dot line shows the signal after glucose was delivered to the assay device.
- the width of color is indicated by the length of the absorption shown by the glucose dot line. Note the variation in diffuse reflectance from A to B is due in part to the baseline variation shown in the red trace. C indicates the approximated width of the assay.
- the width can be converted into number of pixels. This assay may be corrected for background, assay width and edge effects to improve the assay statistics. According to the present invention, three methods are available for evaluating the CMOS data. The data obtained from these algorithms correlate with glucose concentration.
- Figure 25 shows an overlay of CMOS glucose assays.
- 300 nanoliters of each glucose concentration was delivered to an assay device.
- the actual glucose concentration is determined by measurement using a Yellow Spring Instrument that is well established as reliable and accurate.
- Each curve in this graph represents the average of two measurements.
- the data in Fig. 26 may be plotted with the logarithm of the glucose concentration that is linear with signal, as illustrated in Fig. 27 .
- the least squares coefficients are determined to evaluate the accuracy of the predicted, least squares value in relation to measured values. These least square coefficients are shown on Fig. 27 for all points and for the set without the "low" point.
- the second exemplary algorithm is based on "peak value" where the minimum value between pixel 450 and pixel 550 was used as our signal; this value is then subtracted from the background. We plot this net value against the glucose concentration as shown in Figure 28 .
- the third exemplary algorithm is based on "peak area". At each pixel we take the difference between the background and the signal and then we sum up this net value from pixel 450 to pixel 550. We plotted this against glucose concentration. The result is shown in Figure 29 . Lastly, Figure 30 is a plot of glucose concentration versus detector signal for four different test volumes illustrating the volume independence advantage of the present invention.
- the following table summarizes the analyzed results for all three methods.
- the "predicted glucose” is the calculated glucose value using logarithmic fit function.
- the "r-square” is the best-fit r square value using logarithmic fit.
- the "error” is the percentage error in compares our calculated glucose with YSI reading.
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Claims (10)
- Anordnung (80), die konstruiert und angeordnet ist, um das Vorliegen oder die Konzentration eines Analyts in einer Körperflüssigkeit (2108) zu detektieren, wobei die Anordnung Folgendes umfasst:ein Kanalelement (82), das ausgebildet ist, um eine Probe der Körperflüssigkeit (2108) aus einer Leitung (14) aufzunehmen;ein Testfeld (20), das sich in Fluidkommunikation mit einer offenen Unterseite des Kanalelements (82) befindet, wobei das Testfeld (20) ein Reagens (1106) enthält;eine Vielzahl von Detektoren (85), wobei jeder Detektor geeignet ist, um ein Signal zu erzeugen, das eine Reaktion zwischen dem Analyt und dem Reagens (1106) anzeigt;Elektronik, die geeignet ist, um die durch die Vielzahl von Detektoren erzeugten Signale zu überwachen und zu deuten, um Folgendes bereitzustellen:(i) eine Schätzung des Volumens der durch das Kanalelement (82) aufgenommenen Probe, zumindest teilweise auf Grundlage der Anzahl der Detektoren (85), die ein Signal erzeugen, das eine Reaktion zwischen dem Analyt und dem Reagens anzeigt;(ii) Informationen über die Kinetik der Reaktion zwischen dem Analyt und dem Reagens, zumindest teilweise auf Grundlage der Zeitabstände zwischen den Signalen.
- Anordnung nach Anspruch 1, worin die Elektronik ferner zumindest einen Korrekturalgorithmus umfasst.
- Anordnung nach Anspruch 1, worin die Elektronik geeignet ist, um ein Gesamtergebnis auf Grundlage der durch die einzelnen Detektoren (85) erzeugten Signale zu erzeugen.
- Anordnung nach Anspruch 1, die ferner ein Hautpenetrationselement (2102) umfasst, worin das Hautpenetrationselement eine Mikronadel umfasst.
- Anordnung nach Anspruch 1, worin die Vielzahl von Detektoren (85) eine Detektoren-Matrix umfasst.
- Verfahren, das nützlich zur Detektion des Vorliegens oder der Konzentration eines Analyts in einer Körperflüssigkeit ist, wobei das Verfahren Folgendes umfasst:das Einbringen einer Probe der Körperflüssigkeit (2108) in ein Kanalelement (82), das die Probe mit einem Testfeld in Kontakt bringt, wobei das Testfeld (20) ein Reagens enthält;das Erzeugen einer Vielzahl von Signalen durch eine Vielzahl von Detektoren (85), wenn eine Reaktion zwischen dem Analyt und dem Reagens stattfindet;das Überwachen und Deuten der Vielzahl von Signalen, um eine der folgenden Ausgaben bereitzustellen:(i) eine Schätzung des Volumens der durch das Kanalelement (82) aufgenommenen Probe, zumindest teilweise auf Grundlage der Anzahl der Detektoren (85), die ein Signal erzeugen, das eine Reaktion zwischen dem Analyt und dem Reagens anzeigt;(ii) Informationen über die Kinetik der Reaktion zwischen dem Analyt und dem Reagens, zumindest teilweise auf Grundlage der Zeitabstände zwischen den Signalen.
- Verfahren nach Anspruch 6, worin (c) ferner das Anwenden zumindest eines Korrekturalgorithmus auf die das zumindest eine Signal umfasst.
- Verfahren nach Anspruch 6, worin (c) ferner das Erzeugen eines Gesamtergebnisses auf Grundlage der durch die Vielzahl von Detektoren (85) erzeugten Signale umfasst.
- Verfahren nach Anspruch 6, das ferner die Bereitstellung eines Hautpenetrationselements (2102) umfasst, wobei das Hautpenetrationselement eine Mikronadel umfasst.
- Verfahren nach Anspruch 6, worin (b) ferner das Erzeugen zumindest eines Signals durch eine Detektoren-Matrix (85) umfasst.
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US10/394,230 US7052652B2 (en) | 2003-03-24 | 2003-03-24 | Analyte concentration detection devices and methods |
US45696103P | 2003-03-25 | 2003-03-25 | |
US45799603P | 2003-03-28 | 2003-03-28 | |
EP04758045.1A EP1625386B8 (de) | 2003-03-24 | 2004-03-24 | Vorrichtungen und verfahren zum nachweis von analytkonzentrationen |
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EP04758045.1 Division | 2004-03-24 |
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-
2003
- 2003-03-24 US US10/394,230 patent/US7052652B2/en not_active Expired - Lifetime
-
2004
- 2004-03-24 EP EP10180848.3A patent/EP2322914B1/de not_active Revoked
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- 2005-05-10 US US11/125,107 patent/US7427377B2/en not_active Expired - Lifetime
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2008
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- 2012-07-30 US US13/562,129 patent/US9095292B2/en not_active Expired - Lifetime
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US8231832B2 (en) | 2012-07-31 |
US7427377B2 (en) | 2008-09-23 |
EP2322914A1 (de) | 2011-05-18 |
US20090054810A1 (en) | 2009-02-26 |
US20050202567A1 (en) | 2005-09-15 |
US20040191119A1 (en) | 2004-09-30 |
US20120296179A1 (en) | 2012-11-22 |
US9095292B2 (en) | 2015-08-04 |
US7052652B2 (en) | 2006-05-30 |
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